A nanoscale carrier protein platform for enhancing antigen immunogenicity
A technology of antigen immunization and carrier protein, which is applied in the direction of carrier-binding antigen/hapten components, antibody medical components, and resistance to vector-borne diseases. It can solve problems such as increasing HDL-C degradation and research in the field of vaccine nanotechnology. Achieve the effect of enhancing immunity and facilitating recognition
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Embodiment 1
[0054] Embodiment one, the preparation of OVA nano material
[0055] The specific method is as follows:
[0056] Step 1. Dissolving OVA molecules in ddH 2 After O, the mass concentration of the solution is 10mg / ml;
[0057] Step 2, take 2.5ml of the above OVA solution in a 5ml reaction bottle (containing a matching magnet), add 2.5ml of Mes (PH=6.0) buffer and mix;
[0058] Step 3. Put the water bath in a magnetic stirrer, heat it to 70°C, adjust the speed to 750r, put the above reaction bottle in the water bath, and let it fully heat for 2min10s;
[0059] Step 4, the product is dialyzed (ddH 2 (O 5L, change the water every 2 hours, 5 times in total), and freeze-dry to obtain OVA nanomaterials (OVA-NP) with an average particle size of 50nm.
[0060] In the preparation process of the OVA nanomaterial, according to the difference of the target particle size, the reaction time, the concentration of the reaction, the rotation speed and the number of dialysis after the post-tre...
Embodiment 2
[0077] Embodiment two, the preparation of nano vaccine
[0078] The specific preparation formula of the nano vaccine is as follows:
[0079]
[0080] Program A.peptide for CETP 471-476 The specific preparation process of the nano vaccine is as follows:
Embodiment A-1
[0082] Step 1, configuration of reaction solvent:
[0083] Reaction solution A: Dissolve OVA-NP in PBS (pH=7.2, 0.01M), and configure a solution with a mass concentration of 2 mg / ml;
[0084] Reaction solution B: Dissolve NEM in PBS (pH=7.2, 0.01M), and prepare a solution with a mass concentration of 0.6256 mg / ml;
[0085] Reaction solution C: dissolve sulfo-smcc in ddH 2 O, configuration mass concentration is the solution of 4.8mg / ml;
[0086] Reaction solution D: CETP 471-476 (Customized synthesis by Jill Biochemical Company, with a cysteine added at the amino terminal) dissolved in DMF or DMSO, and a solution with a mass concentration of 25mg / ml was prepared;
[0087] Step 2, take 2ml of solution A and 1.078ml of solution B, and react for 2 hours at room temperature at a speed of 400r / min;
[0088] Step 3: Add 0.164ml of solution C, and react for 1 hour at room temperature at a speed of 400r / min;
[0089] Step 4, use 3K ultrafiltration tube, secondary water, 4500r*20...
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